Heavy metal accumulation in Pelargonium hortorum

Heavy metal accumulation in Pelargonium hortorum:
Effects on growth and development
Acumulación de metales pesados en Pelargonium hortorum:
Efectos sobre el crecimiento y el desarrollo
Orroño DI & RS Lavado*
Abstract. Ornamental plants have been proposed for growing in
heavy metal (HM) contaminated soils, and also for phytoremediation. We evaluated (1) biomass production and (2) HM accumulation in Pelargonium hortorum. Plants were grown for 16 weeks on
HM (cadmium, chromium, copper, lead, nickel and zinc) enriched
soils. Treatments were i) control, non-enriched soil, ii) medium concentration treatment, and iii) high concentration treatment. Four
destructive harvests were carried out, and roots, stems, leaves, and
flowers were analyzed each time. Concentrations of HM were determined using ICP. Significant reductions in biomass production were
observed in HM-enriched soils compared with the control. Different indexes confirmed that P. hortorum was affected by HM. Heavy
metals concentrations were higher in roots than shoots. Plant uptake
rates of HM in roots and shoots showed different patterns for each
element. Flowering was highly sensitive to soil HM concentrations.
Key words: Heavy metals, Ornamental plants, Tolerance index,
Enriched soils, Plant heavy metal accumulation.
Resumen. Las plantas ornamentales se han propuesto para crecimiento en suelos contaminados con metales pesados, y también para
su uso en fitoremediación. Se evaluó la producción de biomasa y la
acumulación de metales pesados en plantas de Pelargonium hortorum.
Las plantas crecieron durantes 16 semanas en suelos enriquecidos con
cadmio, cromo, cobre, plomo, níquel y cinc. Los tratamientos fueron
i) control, suelo no enriquecido, ii) tratamiento concentración media y
iii) tratamiento concentración alta. Se realizaron cuatro cosechas destructivas y se analizaron raíces, tallos, hojas y flores en cada cosecha.
Las concentraciones de metales pesados se determinaron mediante
espectrometría de emisión óptica con plasma acoplado inductivamente. La producción de biomasa fue reducida significativamente en los
suelos enriquecidos con metales pesados comparado con las plantas
en suelo control. Diferentes índices confirmaron que P. hortorum fue
afectada negativamente por la aplicación de metales pesados. Las concentraciones de metales pesados fueron mayores en las raíces que en la
parte aérea de las plantas. La tasa de absorción radical y la producción
de biomasa mostraron diferentes patrones dependiendo de los metales
pesados estudiados. La floración fue un parámetro muy sensible a la
concentración de metales pesados en el suelo.
Palabras clave: metales pesados, plantas ornamentales, índice de
tolerancia, suelos enriquecidos, acumulación de metales pesados en
planta.
Facultad de Agronomía, Universidad de Buenos Aires. Av. San Martín 4453
C1417DSE Buenos Aires, Argentina.
* Address Correspondence to: Raúl S. Lavado. Facultad de Agronomía, Universidad de Buenos Aires. Av. San Martín 4453 C1417DSE Buenos Aires, Argentina. e-mail: [email protected]
Recibido/Received 4.VI.2009. Aceptado/Accepted 18.VI.2009.
FYTON ISSN 0031 9457, (2009) 78: 75-82
76
Orroño DI & RS Lavado, FYTON 78 (2009)
INTRODUCTION
Several ornamental plants have been proposed suitable
for growing in heavy metal (HM) contaminated soils, and
also phytoremediation. KrishnaRaj et al. (2000) assessed the
capacity of scented geranium (Pelargonium sp. ‘Frensham’) to
tolerate and accumulate Cr, Ni, and Pb in a hydroponics system. These data provided quantitative evidence for HM tolerance and accumulation potential in the genus Pelargonium.
Orroño et al. (2008) found that Pelargonium hortorum showed
better tolerance to HM than other Pelargonium species. Occurrence of heavy metals in soils is a consequence of industrialization, particularly in soils of urban areas which show
higher levels of HM than those in rural regions (Kelly et al.,
1996; Lavado et al., 1998).
Heavy metal accumulation in plants has received much
attention, and those above-cited studies can be viewed from
three different points of view. Firstly, the effect of HM on
plant biomass production and HM accumulation. When toxic
elements are absorbed by plants, toxicity problems (at a biochemical level) usually result in reduced biomass production
either at a plant organ or whole plant scale (Hagemeyer, 1999).
Decreases in plant biomass are generally correlated with an
increase in absorbed elements concentration (Marschner,
1995). For instance, high metal concentrations were measured
on shoots following growth inhibition of plants subjected to
metal toxicity (Hajiboland, 2005). Therefore, any plant growing in HM contaminated soil usually increases their HM tissue concentration. Heavy metal uptake rates (UR) were developed to quantify HM transport from soil to plants (Singh
& Agrawal, 2007).
Secondly, the timing when plants accumulate HM. Most
research has developed short-term studies. For example, Lutts
et al. (2004) studied the accumulation of cadmium and zinc
during a few days after germination of Atriplex halimus. Few
studies have addressed how HM concentrations vary at different plant growth stages. Perronnet et al. (2003) reported that
cadmium and zinc accumulation varied with plant organs and
age during growth of the hyperaccumulator Thlaspi caerulescens. Stem zinc concentration decreased with time, while cadmium concentration remained constant, despite an increase
in biomass. Young leaves exhibited higher cadmium concentration than older leaves, but the reverse was true for zinc.
Dinelli & Lombini (1996) found that metal concentrations
in plant organs were higher at early vegetative growth stages
due to relatively higher nutrient uptake than plant growth rate
on soils derived from copper mine spoils. At flowering time,
minimal copper and other heavy metal concentrations were
detected. Other works have shown that metals may influence
resource allocation during sexual reproduction (Saikkonen et
al., 1998), and delay flowering (Brun et al., 2003).
Lastly, plant organs differ in HM accumulation. In most
plants, roots, stems, leaves, fruits and seeds exhibit different
FYTON ISSN 0031 9457, (2009) 78: 75-82
HM concentrations, with roots containing the highest and
seeds the lowest HM levels (Kloke et al., 1994). Different indexes have been proposed to evaluate the capacity of a plant
for metal accumulation in harvested organs. This is useful for
phytoremediation. Among those indexes are the shoot:root
ratio and the tolerance index (TIN) (Antosiewicz, 1995). The
relative growth rate (RGR) is also used to determine the ability of a plant to accumulate metals (Lutts et al., 2004). The
first two indexes, are usually applied at final harvest, but the
latter needs to be applied at different harvesting times during
the growing season.
We studied the ability of the ornamental plant species Pelargonium hortorum (Geraniaceae), ‘Common Geranium’, for
growing on HM contaminated soils. Our objectives were i)
to determine the accumulation and its uptake rate in roots,
stems, leaves and flowers of HM, throughout different harvesting times; ii) to evaluate biomass production by applying
HM accumulation indexes and iii) to analyze the effect of
HM on flowering development.
MATERIALS AND METHODS
We conducted a pot study using a Typic Argiudoll soil.
Physical and chemical properties of this soil were measured
using standard techniques (Klute, 1986; Sparks et al., 1996);
major values for these properties were: sand 17.1%; silt 57.1%;
clay 25.8%; 1: 2.5 (w/v) pH 6.1; organic matter 3.90%; total nitrogen (Kjeldahl) 0.22%; extractable phosphorus (Bray
& Kurtz) 12.5 mg/kg; and exchangeable potassium 1.35
meq/100g. A completely randomized design with three treatments and eight replications per treatment were performed.
Treatments were as follows: i) Control, not contaminated
soil (T0), ii) Medium HM concentration (T1), and iii) High
HM concentration (T2). Treatments T1 and T2 received cadmium (Cd) as cadmium nitrate; chromium (Cr) as chromic
acid; copper (Cu) as copper chloride; lead (Pb) as lead nitrate;
nickel (Ni) as nickel sulfate, and zinc (Zn) as zinc sulfate. The
T1 treatment contained the following HM concentrations:
10, 250, 250, 80, 500 and 300 mg/kg Cd, Cu, Cr, Ni, Pb and
Zn, respectively. The T2 treatment included the same metals,
but twice as much the T1 concentration treatment. Salts were
carefully mixed with the soil, and subjected to wet/dry cycles
during a three month period prior to the onset of the experiment. This procedure reduced any overestimation of metal
bioavailability (Basta et al., 2005). It was because after mixing,
metals interacted with the soil matrix reaching a new equilibrium (Martinez & Motto, 2000).
Uniform plantlets were selected, and three were transplanted to plastic pots of 2000 cm3 volume. Subsequently, only
one plant per pot was maintained, and the other two were
removed. Approximately every four weeks, over a period of 16
weeks, two plants were harvested from each treatment. Plant
material was oven-dried at 60°C after washing to determine
Heavy metals on Pelargonium hortorum
dry weight and heavy metal content in roots, stems, leaves,
and flowers. Plant material was first digested with a nitricperchloric acid mixture ( Jones & Case, 1990), and HM content was then determined by ICP-AES (Sparks et al., 1996),
and expressed on a dry weight basis.
Several indexes were calculated: (1) Shoot/root ratios; (2)
TIN, which represents the biomass ratio for plants grown in
HM enriched-soils relative to those grown in HM non-enriched, control-soils (Antosiewicz, 1995); (3) the RGR for
each harvesting interval, based on the following equation:
RGR = (ln W2 - ln W1) / (t2 - t1),
where W2 and W1 represent total plant dry weights at
times t1 and t2, respectively (Lutts et al., 2004).
Finally, UR (mg.plant-1.d-1) were calculated as:
Heavy metal uptake rates (UR) = M2*W2 – M1*W1
t2 – t1
where M1 and M2 are metal concentrations in plant tissue,
and W1 and W2 are plant biomasses at times t1 and t2 (Singh
& Agrawal, 2007).
Dry matter yields, HM concentrations in different organs,
shoot/root ratios, and RGR were analyzed using Statistics 8.0.
Data were subjected to a two-way analysis of variance (ANOVA) for each sampling time to examine significant differences
between treatments (T0, T1, and T2), harvesting times, and
HM concentrations in different plant organs. Differences between individual means were tested using Tukey’s Test at the
0.05 significance level.
RESULTS
Heavy metal stress had no effect on plant survival, and all
plants were alive at the end of the experiment. However, plant
biomass was affected by increases in soil HM concentrations
(Table 1). Aerial biomass production was significantly reduced
(p<0.05) at 3rd and 4th harvests in plants from the T1 and T2
treatments compared to T0 plants. At the last sampling date,
biomass was lower (p<0.05) in the T2 treatment for stems,
and in T1 treatment for leaves, compared to T0 treatments.
The interaction of treatment x harvest was significant (p<0.05).
In T1 treatments some replications did not produce flowers,
while in T2 treatments there was not flower production at
all. Root biomass decreased significantly (p<0.05) in T2 treatments compared with the T0 and T1, but there were no differences (p>0.05) among harvesting times within each treatment (Table 1).
A change in biomass distribution between roots and shoots,
and the inhibition of shoot growth on HM contaminated soils
was confirmed by the shoot/root ratio. In T0 treatment this
ratio increased significantly from 7 to 16 from the first to the
fourth harvest. Meanwhile, the shoot/root ratio in both the T1
77
Table 1. Biomass accumulation in Pelargonium hortorum per treatment and harvesting time.
Tabla 1. Acumulación de biomasa en Pelargonium hortorum por tratamiento y fecha de cosecha.
Harvest
1
2nd
3
Treat.
st
rd
T0
4th
2
3rd
T1
4
th
1st
2
nd
3rd
4
th
T2
Stems
Leaves
Flowers
0.6 ± 0.1 a 2.1 ± 0.3 b 2.3 ± 0.2 abc
0.8 ± 0.9
0.6 ± 0.3 a 3.5 ± 0.5 ab
1.5 ± 0.4
0.4 ± 0.1 a 1.7 ± 1.0 b
0.8 ± 0.1 a
1st
nd
Roots
Biomass (g)
4.2 ± 0.3 a
0.6 ± 0.2 c
4.2 ± 0.7 ab
5.4 ± 1.22 a
0.6 ± 0.1
1.3 ± 0.5
0.5 ± 0.2 b 2.1 ± 0.1 b 1.9 ± 0.9 bc
0.3*
0.4 ± 0.1 a 1.8 ± 0.4 b 1.2 ± 1.3 bc
0.5*
0.6 ± 0.3 a
2.2 ± 0.4 ab
1.4 ± 1.0 bc
0.9 ± 0.1 a 2.3 ± 0.3 ab
0.6 ± 0.0 c
0.4*
0.2*
0.5 ± 0.0 b 2.1 ± 0.3 b 0.7 ± 0.1 bc
nfp
0.4 ± 0.2 b 2.5 ± 0.7 ab 0.1 ± 0.1 ab
nfp
0.4 ± 0.1 b 1.5 ± 0.6 b 0.4 ± 0.0 abc
0.3 ± 0.1 b 1.9 ± 0.2 b 0.2 ± 0.2 bc
nfp
nfp
*no replicates
*falta de réplicas
nfp: no flower production
nfp: no producción de flores
Means followed by the same letter do not differ statistically at p≤0.05.
Promedios seguidos por la misma letra no difieren significativamente a p≤0,05.
and T2 treatments remained between 4 to 8, and it was significantly lower (p<0.005) in T2 treatment (data not shown).
The tolerance index showed significant differences between
harvesting times (p<0.05) (Fig. 1), and T1 treatment exhibited
TIN values greater than 100% for the 1st and 2nd harvest, indicating a net increase in biomass relative to the control. TIN
values for T1 and T2 treatments were lower than 100% for the
3rd and 4th harvests, showing a net decrease in biomass, and
growth inhibition. Relative growth rates increased throughout
the experiment (Fig. 2). However, they were lower (p<0.05) in
HM contaminated soils (T2) than in the control, showing a
delay in plant growth rates.
In general, the increase in HM content in plants was
consistent with the metal concentration in soil. Heavy metal
accumulation in roots (Table 2) was higher than that in the
other organs (roots > stems > leaves > flowers). Only Cu and
Zn were detectable in all treatments, but they showed no significant accumulation differences among the T0, T1 and T2
throughout the study. The other HM´s were not detectable
in the Control. Heavy metal accumulation in T1 treatment,
at different harvesting times (Table 2) showed a similar pattern to T2 treatment. Root Cd, Ni, and Zn accumulations
increased significantly over time, and continued to accumulate throughout the experiment. Chromium, Cu and Pb, on
the other hand, exhibited no consistent trend throughout
time. Accumulation of HM in stems and leaves (Tables 3
FYTON ISSN 0031 9457, (2009) 78: 75-82
78
Orroño DI & RS Lavado, FYTON 78 (2009)
Fig. 1. Tolerance index of Pelargonium hortorum plants for Medium
(T1) and High HM concentration (T2) treatments for different harvesting dates. Means followed by the same letter do no differ statistically
(p≤0.05; Tukey´s test).
Fig 1. Índice de tolerancia de las plantas de Pelargonium hortorum en los
tratamientos concentración media (T1) y alta (T2) en diferentes fechas de
cosecha. Las medias seguidas por la misma letra no difieren estadísticamente (p≤0,05; Tukey´s test).
160
a
140
T1
a
80
a
60
T2
a
b
40
20
0
b
b
st
nd
1 harvest
b
rd
2 harvest
th
3 harvest
4 harvest
Fig. 2. Relative growth rate (RGR) of Pelargonium hortorum plants
during the experimental period. Means followed by the same letter do
no differ statistically (p≤0.05; Tukey´s test).
Fig 2. Tasa relativa de crecimiento (TRC) de la plantas de Pelargonium
hortorum durante el periodo experimental. Las medias seguidas por la
misma letra no difieren estadísticamente (p≤0,05; Tukey´s test).
0.005
RGR (g/g dry matter/d)
0.000
-0.005
30
60
90
b
-0.030
-0.035
T0
T1
Time (days)
and 4) showed no significant differences (p<0.05); although a
subtle pattern was inferred as plant growth proceeded, leaves
at the 4th harvesting time exhibited lower HM levels than
leaves at the 1st harvesting time. There was a significant treatment effect (p<0.05), but differences in HM concentrations
for harvesting times were not significant (p>0.05) in stems
and leaves. The exceptions were HM concentrations of Cd, Ni
and Pb in plant roots (Table 2). This result was likely due to a
consistently higher concentration of HM in roots. A significant treatment x harvest interaction (p<0.05) was detected for
root Cd and Ni, but in general the accumulation of HM did
not increase considerably over the growing season.
Table 5 summarizes flower HM concentrations among the
three treatments. Only Cu and Zn were measured in the ConFYTON ISSN 0031 9457, (2009) 78: 75-82
T0
Zn
0.250
T2
b
Cu
0.300
b
a
a
b
ab
ab
-0.015
-0.025
120
ab
-0.010
-0.020
Fig. 3. Tasa de absorción de metales pesados (mg/plant/d) de raíces (a)
y biomasa aérea (b) en plantas de Pelargonium hortorum en el tratamiento
control en función de la fecha de cosecha.
a
a
Fig. 3 Heavy metal uptake rate (mg/plant/d) for roots (a) and aerial
biomass (b) of Pelargonium hortorum in T0 treatment as a function of
harvesting date.
UR (mg/plant/d)
100
0.200
0.150
0.100
0.050
0.000
-0.050
30
-0.100
0.100
60
90
Time (days)
Cu
Zn
120
a
T0
0.080
UR (mg/plant/d)
TIN (%)
120
trol. Although T1 treatment did not have an adequate number
of flowers for statistical analysis, it appears that HM exhibited
an increase in concentrations with respect to T0 treatment.
The high HM concentrations in T2 resulted in the absence
of flowering.
Figure 3 shows Cu and Zn uptake rates (mg/plant/d) for
roots and aerial biomass in T0 treatment. Rates of Cu uptake
by plants were very stable for roots and aerial biomass. Zn
showed variable uptake rates, especially in roots, but there was
not a clear tendency across the duration of the study. Zinc uptake rates by roots in T1 treatment (Fig. 4) were positive at the
beginning and end of the growth period, and the lower uptake
rates occurred at 90 days from study initiation. Cadmium, Cu
and Pb, and to a lesser extent Ni, uptake rates were basically
constant throughout harvests. Rates of Zn uptake in aerial
biomass were positive in T1 treatment (Fig. 4). Zinc, and to a
lesser extent Ni, tended to decrease across time. The remaining
HM showed more stable uptake rates. Uptake rates of HM by
roots and the aerial part were less clear at T2 treatment (Fig. 5).
These plant organs showed either positive Zn and Ni uptake
rates at the beginning or negative uptake rates for these HM
at the end of the growth period. Rate of uptake progressively
decreased with plant development. Cadmium, Cr, Cu and Pb
were more stable.
0.060
0.040
0.020
0.000
-0.020
30
60
90
Time (days)
120
b
Heavy metals on Pelargonium hortorum
79
Table 2. HM accumulation in roots of Pelargonium hortorum per treatment and harvesting date.
Tabla 2. Acumulación de metales pesados en raíces de Pelargonium hortorum por tratamiento y fecha de cosecha.
Harvest
Treatm
1st
2
nd
3
rd
T0
1
3rd
T1
4
1
3
4
bdl
11.4 ± 1.5
323.7 ± 36.5
122.4 ± 43.3
646.8 ± 65.8
T2
th
ANOVA
77.8 ± 17.1
118.7 ± 8.0
st
rd
84.3 ± 60.1
187.0 ± 58.0
th
2nd
Cd
6.3 ± 0.4
st
2
Zn
12.9 ± 2.9
4th
nd
Cu
Treatm.
748.2 ± 398.2
466.1 ± 125.3
941.8 ± 80.6
***
Harvest
50.5 ± 145.5
53.8 ± 4.2
26.6 ± 0.9
bdl
bdl
40.8 ± 3.2
3815.9 ± 22.4
40.2 ± 1.4
25.8 ± 0.6
5003.2 ± 2127.1 73.2 ± 10.4
7271.7 ± 932.9 94.4 ± 13.9
9501.9 ± 966.0 129.0 ± 10.7
9417.1 ± 699.1 120.5 ± 12.8
***
***
ns
*
ns
ns
Cr
Pb
bdl
bdl
bdl
bdl
bdl
bdl
3596.3 ± 4.9
2753.7 ± 17.5
Ni
bdl
2397.9 ± 283.7 24.6 ± 3.2
ns
Treat x harv.
Metal in roots (ppm)
bdl
bdl
bdl
323.3 ± 14.4
473.3 ± 64.4
359.8 ± 14.2
721.5 ± 90.7
815.6 ± 169.9
1009.1 ± 3.2
1245.3 ± 16.9
bdl
bdl
100.7 ± 37.3
123.8 ± 24.1
89.6 ± 14.8
90.5 ± 22.5
225.8 ± 34.6
347.5 ± 89.2
194.8 ± 19.4
1550.5 ± 228.0 353.0 ± 11.5
*
**
***
*
ns
**
bdl
164.3 ± 59.3
222.1 ± 22.6
99.5 ± 44.6
111.7 ± 12.7
202.4 ± 46.4
340.5 ± 52.8
152.2 ± 34.0
270.1 ± 45.6
***
ns
*
ns
Significance levels for treatments, harvesting times and the interaction treatment × harvesting times are shown: *p<0.05; **p<0.01; ***p<0.001; bdl:
Below detection limit.
Niveles de significancia para tratamientos, épocas de cosecha, y la interacción tratamiento x época de cosecha: *p<0,05; **p<0,01; ***p<0,001; bdl: Por debajo
del límite de detección.
Table 3. HM accumulation in stems of Pelargonium hortorum per treatment and harvesting time.
Tabla 3. Acumulación de metales pesados en tallos de Pelargonium hortorum por tratamiento y fecha de cosecha.
Harvest
Treatm
1st
2
nd
3
rd
T0
1
3rd
T1
4
1
3
4
bdl
4.4 ± 0.2
24.8 ± 4.9
37.4 ± 9.5
104.7 ± 47.6
T2
th
ANOVA
27.9 ± 8.0
33.2 ± 4.4
st
rd
3.5 ± 1.5
26.2 ± 0.6
th
2nd
Cd
1.9 ± 1.9
st
2
Zn
3.25 ± 2.7
4th
nd
Cu
Treatm.
Harvest
Treat x harv.
103.7 ± 23.9
57.2 ± 47.5
51.8 ± 3.2
***
ns
ns
43.0 ± 8.1
41.7 ± 12.0
14.9 ± 12.9
449.2 ± 42.3
672.2 ± 195.9
1010.6 ± 66.6
1110.7 ± 101.1
Metal in stems (ppm)
bdl
bdl
7.8 ± 0.8
12.6 ± 4.3
14.1 ± 2.2
12.7 ± 2.8
3260.7 ± 3155.9 38.0 ± 33.1
3333.2 ± 417.9 37.4 ± 0.1
ns
Pb
bdl
bdl
bdl
bdl
bdl
bdl
5614.5 ± 964.7 96.7 ± 28.9
ns
Cr
bdl
3633.9 ± 1587.3 55.3 ± 29.8
***
Ni
*
ns
ns
bdl
bdl
bdl
83.0 ± 1.6
118.6 ± 39.2
149.9 ± 19.7
166.6 ± 16.1
571.8 ± 243.1
bdl
bdl
20.8 ± 5.1
18.6 ± 3.1
30.4 ± 8.5
26.3 ± 0.6
61.2 ± 33.7
1017.2 ± 179.0 63.8 ± 20.0
830.5 ± 817.2
545.1 ± 15.7
**
ns
ns
42.9 ± 33.1
31.6 ± 0.2
***
ns
ns
bdl
39.1 ± 5.7
43.6 ± 5.8
70.3 ± 18.4
79.9 ± 4.7
113.6 ± 60.5
152.6 ± 48.9
82.8 ± 73.2
74.7 ± 3.7
***
ns
ns
Significance levels for treatments, harvesting times and the interaction treatment × harvesting times are shown: *p<0.05; **p<0.01; ***p<0.001; bdl:
Below detection limit.
Niveles de significancia para tratamientos, épocas de cosecha, y la interacción tratamiento x época de cosecha: *p<0,05; **p<0,01; ***p<0,001; bdl: Por debajo
del límite de detección.
FYTON ISSN 0031 9457, (2009) 78: 75-82
80
Orroño DI & RS Lavado, FYTON 78 (2009)
Table 4. HM accumulation in leaves of Pelargonium hortorum per treatment and harvesting time.
Tabla 4. Acumulación de metales pesados en hojas de Pelargonium hortorum por tratamiento y fecha de cosecha.
Harvest
Treatm
1
Cu
Zn
Cd
6.6 ± 0.7
38.8 ± 0.2
bdl
10.0 ± 0.8
st
2nd
T0
3
rd
50.1 ± 9.4
6.9 ± 1.3
4
36.9 ± 2.4
4.1 ± 0.1
th
1st
26.8 ± 1.1
21.0 ± 5.0
2
nd
15.9 ± 9.9
T1
3
rd
307.9 ± 92.3
300.1 ± 176.0
18.9 ± 2.9
4th
642.5 ± 16.0
7.9 ± 0.1
1
342.6 ± 2.6
32.9 ± 15.0
st
2
nd
48.0 ± 24.1
T2
3rd
4
890.4 ± 15.0
20.1 ± 1.9
Treatm.
643.5 ± 326.3
*
Harvest
***
ns
Treat x harv..
bdl
ns
ns
ns
Ni
Cr
Pb
bdl
bdl
bdl
bdl
bdl
bdl
bdl
bdl
38.0 ± 18.1
75.9 ± 45.1
9.7 ± 0.7
33.3 ± 26.3
148.2 ± 11.4
5.9 ± 0.1
26.0 ± 14.0
17.8 ± 1.9
10.8 ± 1.7
35.8 ± 3.9
137.4 ± 15.5
7.9 ± 0.1
328.0 ± 151.2
46.9 ± 14.9
349.9 ± 130.3
97.0 ± 53.1
160.5 ± 30.6
15.8 ± 0.1
109.4 ± 9.4
***
13.8 ± 4.4
*
ns
*
ns
ns
bdl
bdl
77.0 ± 19.1
8.1 ± 0.0
bdl
bdl
bdl
5.0 ± 2.0
1409.5 ± 711.9 24.0 ± 10.0
13.9 ± 0.1
th
ANOVA
1426.9 ± 689.3
Metal in leaves (ppm)
bdl
59.0 ± 29.1
43.2 ± 36.3
46.7 ± 36.3
17.9 ± 2.2
74.9 ± 28.9
135.0 ± 79.2
21.8 ± 0.2
21.3 ± 2.5
*
ns
ns
ns
ns
ns
Significance levels for treatments, harvesting times and the interaction treatment × harvesting times are shown: *p<0.05; **p<0.01; ***p<0.001; bdl:
Below detection limit.
Niveles de significancia para tratamientos, épocas de cosecha, y la interacción tratamiento x época de cosecha: *p<0,05; **p<0,01; ***p<0,001; bdl: Por debajo
del límite de detección.
Fig. 4. Heavy metal uptake rate (mg/plant/d) for roots (a) and aerial
biomass (b) of Pelargonium hortorum in the Medium HM concentration
treatment (T1) as a function of harvesting date.
Fig. 4. Tasa de absorción de metales pesados (mg/plant/d) de raíces (a)
y biomasa aérea (b) en plantas de Pelargonium hortorum en el tratamiento
concentración media (T1) en función de la fecha de cosecha.
0.100
Zn
Pbx2
Crx2
Cdx2
Nix2
0.040
0.020
0.000
30
60
-0.040
90
120
a
Time (days)
0.100
Cux2
Zn
Pbx2
Crx2
Cdx2
Ni
T1
0.060
0.040
0.020
0.000
-0.020
0.100
0.080
Cux2
Zn
Pbx2
Crx2
Cdx2
Nix2
T2
0.060
0.040
0.020
0.000
-0.020
30
60
-0.040
0.300
0.250
0.200
0.150
0.100
0.050
0.000
-0.050
-0.100
90
120
a
Time (days)
Cux2
Zn
Pbx2
Crx2
Cdx2
Ni
T2
UR (mg/plant/d)
0.080
UR (mg/plant/d)
T1
0.060
-0.020
Fig. 5. Tasa de absorción de metales pesados (mg/plant/d) de raíces (a)
y biomasa aérea (b) en plantas de Pelargonium hortorum en el tratamiento
concentración alta (T2) en función de la fecha de cosecha.
UR (mg/plant/d)
UR (mg/plant/d)
0.080
Cux2
Fig. 5. Heavy metal uptake rate (mg/plant/d) for roots (a) and aerial
biomass (b) of Pelargonium hortorum in the High HM concentration
treatment (T2) as a function of harvesting date.
30
60
FYTON ISSN 0031 9457, (2009) 78: 75-82
Time (days)
90
120
b
30
60
90
Time (days)
120
b
Heavy metals on Pelargonium hortorum
81
DISCUSSION
Aerial growth was affected by HM toxicity more than root
growth, and roots of Pelargonium plants accumulated a significantly greater heavy metal concentration than aerial organs.
Other authors have shown that growth of the upper plant parts
is more sensitive to heavy metals, in spite of their low metal
content compared with roots. It was hypothesized that roots
could play an important role in metal retention by preventing
an excessive and toxic accumulation in shoots (Mazhoudi et al.,
1997). Initial leaf mortality and leaf turnover at harvesting time
were higher at T2 than at the T1 treatment. The accelerated
senescence caused by HM comes from oxidative damage and
increased membrane permeability (Luna et al., 1994).
The lower shoot/root ratios found in plants of T1 and T2
treatments, compared with T0 treatment, clearly show that
biomass allocation changes under heavy metal stress conditions between roots and shoots in Pelargonium plants. Tolerance indexes have been useful to characterize plant tolerance.
TIN values lower than 100% indicate a net decrease in biomass and suggest that plants are HM-stressed. At the same
time, TIN values equal to 100% indicate no differences relative to non-HM control treatments. Also, TIN values greater
than 100% indicate a net increase in biomass, and suggest
that plants express a growth dilution effect (Audet & Charest,
2007). Our TIN values decreased as soil HM concentrations
increased (Fig. 1). RGR values progressively increased for all
treatments; this response in T1 and T2 treatments suggest that
these plants recovered after an initial shock, and that growth
was delayed under HM stress. Negative values in T1 and T2
treatments (Fig. 2) can be explained by increases in senesced
leaf number (Davidson & Campbell 1984); the lower RGR
value in T2 treatment may be a consequence of decreases in
shoot biomass (Table 1).
Low transport of HM to shoots may be due to saturation
of root metal uptake, when internal metal concentrations are
high (Zhao et al, 2003). Although we did not find significant
effects of harvesting times for leaves, new leaves tended to
have lower HM levels than older ones (Table 4). Metal concentrations, therefore, may rise as leaves age simply due to the
continued passive metal transport into leaf tissues. Movement
of metals into older leaves is a way that some plants have to
eliminate some of their metal excess (Verkleij & Schat, 1990).
This result disagrees with that of Marschner (1995), who attributed a decline in dry matter mineral content as plants age
to an increase in the proportion of structural material (cell
wall and lignin) and storage compounds.
By the time of the final harvest, very few plants were flowering on the HM contaminated treatments (Table 5). A delay
and inhibition of flowering can be attributed to a disruption in
biological processes due to HM stress (Van Assce & Clijsters,
1990). Our results suggest that flowering was very sensitive to
HM concentrations (Saikkonen et al., 1998).
Table 5. HM in flowers at different harvesting times. Lack of standard deviations indicates lack of replicates.
Tabla 5. Acumulación de metales pesados en flores en distintas fechas de cosecha. La ausencia de desviación estándar indica ausencia de réplicas.
Harvest
Treatm
1
2nd
3
rd
T0
4th
1
2
3rd
T1
4
1
3
4
10.0 ± 0.0
41.5 ± 12.0
bdl
9.9 ± 0.0
96.8 ± 116.0
13.7
17.8
nfp
st
rd
Cd
16.5
th
2nd
Zn
2.9 ± 4.0
st
nd
Cu
21.2 ± 16.0
st
T2
th
bdl: Below detection limit.
nfp: No flower production.
nfp
13.2
nfp
33.0 ± 11.0
43.6 ± 19.7
24.9 ± 1.2
99.3
139.2 ± 84.7
87.6
333.3
nfp
nfp
153.7
nfp
Metal in flowers (ppm)
bdl
bdl
bdl
16.6
8.4 ± 8.8
2.4
4.4
nfp
nfp
44.1
nfp
Ni
Cr
Pb
bdl
bdl
bdl
bdl
bdl
bdl
15.6
45.3 ± 43.3
87.6
88.9
nfp
nfp
125.2
nfp
bdl
bdl
bdl
16.6
11.1 ± 5.0
5.7
8.9
nfp
nfp
5.9
nfp
bdl
bdl
bdl
15.5
17.1 ± 3.4
8.8
17.8
nfp
nfp
8.3
nfp
bdl: Por debajo del límite de detección.
nfp: No hubo producción de flores.
FYTON ISSN 0031 9457, (2009) 78: 75-82
82
Orroño DI & RS Lavado, FYTON 78 (2009)
REFERENCES
Antosiewicz, D. M. (1995). The relationships between constitutional
and inducible Pb-tolerance and tolerance to mineral deficits in
Biscutella laevigata and Silene inflate. Environmental and Experimental Botany 35: 55–69.
Audet, P. & C. Charest (2007). Heavy metal phytoremediation from
a meta-analytical perspective. Environmental Pollution 147: 231237.
Basta, N.T., J.A. Ryan & R.L. Chaney (2005). Trace element chemistry in residual-treated soil: Key concepts and metal bioavailability. Journal of Environmental Quality 34: 49-63.
Brun, L.A, J. Le Corff & J. Maillet (2003). Effects of elevated soil
copper on phenology, growth and reproduction of five ruderal
plant species. Environmental Pollution 122: 361–368.
Davison, H.R. & C.A. Campbell (1984). Growth rates, harvest index and moisture use of Manitou spring wheat as influenced by
nitrogen, temperature and moisture. Canadian Journal of Plant
Science 64: 825-39.
Dinelli, E. & A. Lomboni (1996). Metal distribution in plants growing on copper mine spoils in Northern Apennines, Italy: the evaluation of seasonal variation. Applied Geochemistry 11: 375-85.
Hagemeyer, J. (1999) Ecophysiology of plant growth under heavy
metal stress. In: Prasad, M.N.V. and Hagemeyer, J. (eds), pp 157181. Heavy Metal Stress in Plants from Molecules to Ecosystem,
Springer-Verlag, Heidelberg,
Hajiboland, R. (2005). An evaluation of the efficiency of cultural
plants to remove heavy metals from growing medium. Plant, Soil
and Environment 51: 156–164.
Jones Jr., J.B. & V.W. Case (1990). Sampling, handling and Analysing Plant Tissue Samples. In: Westerman, R.L. (ed), P. 784.
Soil Testing and Plant Analysis, SSSA Book Series, N° 3.
Wisconsin,USA.
Kelly, J., I. Thornton & P.R. Simpson (1996). Urban geochemistry:
a study of the influence of anthropogenic activity on the heavy
metal content of soils in traditionally industrial and non-industrial areas of Britain. Applied Geochemistry 11: 363–370.
Kloke, A., D.R. Sauerbeck & H. Vetter (1994). Study of the Transfer Coefficient of Cadmium and Lead in Ryegrass and Lettuce.
In: Nriagu, J (ed), p 113. Changing Metal Cycles and Human
Health, Springer Verlag, Berlin.
Klute, A. (1986). Methods of Soil Analysis, Part 1. Physical and
Mineralogical Methods. Soil Science Society of America, Inc.
Madison, WI.
Krishnaraj, S., T.V. Dan & P.K. Saxena (2000). A fragrant solution
to soil remediation. International Journal of Phytoremediation 2:
117-132.
Lavado, R.S., M.G. Rodriguez, J.D. Scheiner, M.A. Taboada, G. Rubio, R. Alvarez, M. Alaconada & M.S. Zubillaga (1998). Heavy
metals in soils of Argentina: Comparison between urban and agricultural soils. Communication in Soil Science and Plant Analysis
29: 1913-1917.
Luna, C.M., C.A. Gonzalez & V.S. Trippi (1994). Oxidative damage
caused by an excess of copper in oat leaves. Plant and Cell Physiology 35: 11-15.
Lutts, S., I. Lefévre, C. Delpérée, S. Kivits, C. Dechamps, A. Robledo
& E. Correal (2004). Heavy Metal Accumulation by the Halophyte Species Mediterranean Saltbush. Journal of Environmental
Quality 33: 1271-1279.
FYTON ISSN 0031 9457, (2009) 78: 75-82
Martinez, C.E. & H.L. Motto (2000). Solubility of Pb, Zn and Cu
added to mineral soils. Environmental Pollution 107: 153-158.
Marschner, H. (1995). Mineral Nutrition of Higher Plants. Academic Press, London, UK.
Mazhoudi, S., A. Chaouhi, M.H. Ghorbal & E. Elferjani (1997).
Response of antioxidant enzymes to excess copper in tomato (Lycopersicon esculentum, Mill.). Plant Science 127: 129-137.
Orroño, D., H.E. Benítez & R.S. Lavado (2009). Effects of heavy
metals in soils on biomass production and plant element accumulation of Pelargonium and Chrysanthemum species. Agrochímica
LIII: 000-000 (In press).
Perronnet, T, K., C. Schwartz & J.L. Morel (2003). Distribution of
cadmium and zinc, in the hyperaccumulator Thlaspi caerulescens
grown on multicontaminated soil. Plant and Soil 249: 19-25.
Saikkonen, K., S. Koivunen, T. Vuorisalot & P. Mutikainen (1998).
Interactive effects of pollination and heavy metals on resource allocation in Potentilla anserina L. Ecology 79: 1620–1629.
Singh, R.P. & M. Agrawal (2007). Effects of sewage sludge amendment on heavy metal accumulation and consequent responses of
Beta vulgaris plants. Chemosphere 67: 2229-2240.
Sparks, D.L., A.L. Page, P.A. Helmke, R.H. Loeppert, P.N. Soltabpour, M.A. Tabatabai, C.T. Johnson & M.E. Summer (1996).
Methods of Soil Analysis. Part 3. Chemical Methods. Soil Science
Society of America, Inc . Madison, WI, USA
Van Assche, F. & H. Clijsters (1990). Effects of heavy metals on enzyme activity in plants. Plant and Cell Environment 13: 195-206.
Verkleij, J.A., & H. Schat (1990). Mechanisms of metal tolerance
in higher plants. In: Shaw. A.J. (ed), pp 179-194. Heavy metal
tolerance in plants: evolutionary aspects, CRC Press. Boca Raton,
FL, USA.
Zhao, F.J., E. Lombi & S.P. McGrath (2003). Assessing the potential
for Zn and cadmium phytoremediation with the hyperaccumulator Thlaspi caerulescens. Plant and Soil 249: 37-43.